Canine parvovirus resistant genetic engineering antibody and application thereof

A technology of genetically engineered antibodies and canine parvovirus, applied in genetic engineering, antiviral agents, antiviral immunoglobulins, etc., can solve problems such as reducing curative effect, and achieve the effect of inhibiting agglutination and good neutralizing activity

Pending Publication Date: 2020-02-28
CHANGCHUN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Currently, the canine parvovirus monoclonal antibody and antiserum on the market have strong immunogenicity, and th

Method used

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  • Canine parvovirus resistant genetic engineering antibody and application thereof
  • Canine parvovirus resistant genetic engineering antibody and application thereof
  • Canine parvovirus resistant genetic engineering antibody and application thereof

Examples

Experimental program
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Effect test

Example Embodiment

[0034] Example 1

[0035] Canineization of murine CPV monoclonal antibody

[0036] (1) Construction of canine CPV genetic engineering antibody

[0037] Using the method of the patent application description example of the publication number CN109627331A, the mouse CPV monoclonal antibody was obtained, and the nucleotide sequence of its heavy chain variable region was sequenced as shown in SEQ ID NO.1, and the nucleotide sequence of its light chain variable region was The nucleotide sequence is shown in SEQ ID NO.2, connected to the cloning vector, the clones were picked for sequencing, and the sequencing results were analyzed online using IMGT to obtain the heavy chain amino acid sequence of the antibody sequence as shown in SEQ ID NO.3, and the light chain The amino acid sequence is shown in SEQ ID NO.4.

[0038] The heavy chain variable region gene sequence of the obtained mouse CPV monoclonal antibody was assembled with the canine antibody heavy chain constant region gene...

Example Embodiment

[0039] Example 2

[0040] Expression and activity of canine CPV genetically engineered antibody

[0041] (1) Eukaryotic expression of canine CPV genetic engineering antibody

[0042] Take 1×10 6 Cells / well cell density HEK-293 adherent cells were seeded in six-well plates and grown for 12 hours before transient transfection experiments. According to light chain:heavy chain = 1:1 mixed with PEI (1 μg / μL) at a ratio of 1:3, it was transfected into HEK-293 cells for expression, and the supernatant was collected 72 hours after transfection for activity identification.

[0043] (2) Hemagglutination inhibition method was used to measure the resistance effect of the canine-derived CPV genetically engineered antibody in the supernatant on the binding of canine parvovirus and porcine red blood cells

[0044] Add 25 μL / well of PBS (0.15M pH6.5) to the 96-well V-type hemagglutination plate, add 25 μL of canine CPV genetic engineering antibody to the first column, and do 3 replicates f...

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Abstract

The invention provides a canine parvovirus resistant genetic engineering antibody and an application thereof, and relates to the technical field of antibody engineering. A variable region sequence ofan acquired canine parvovirus monoclonal antibody is assembled with a canine antibody constant region to obtain the canine parvovirus resistant genetic engineering chimeric antibody, according to experimental verification, the hemagglutination inhibition titer of cell supernatant of the genetic engineering antibody is 1:(24-25), and the neutralizing titer of the cell supernatant is 1:203. Obviously, the genetic engineering antibody shows good canine parvovirus neutralizing activity, agglutination of red blood cells by canine parvoviruses can be inhibited, and the canine parvovirus resistant genetic engineering antibody can be applied to the field of canine research of the monoclonal antibody, canine parvovirus control and the like, and is of great significance for promoting development ofcanine monoclonal antibody drugs.

Description

technical field [0001] The invention belongs to the technical field of antibody engineering, and in particular relates to a genetically engineered antibody against canine parvovirus and its application. Background technique [0002] Canine parovirus (canine parovirus, CPV) belongs to the Parvovirus genus of the family Parvoviridae, and is an important pathogen of domestic dogs and several wild carnivores. The main natural host is dogs. Infected dogs can cause canine parvovirus disease, which is divided into enteritis type and myocarditis type. It has the characteristics of high incidence, strong infectivity, and high mortality. It can cause serious economic losses and seriously endanger the development of China's dog breeding industry. [0003] Vaccines to prevent canine parvovirus mainly include: inactivated vaccines, attenuated vaccines and multi-linked vaccines, but research on the immune effects of vaccines on the market has found that only 20% of vaccines can produce 10...

Claims

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Application Information

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IPC IPC(8): C07K16/08C12N15/13A61K39/42A61P31/20
CPCC07K16/081A61P31/20C07K2317/24C07K2317/76A61K2039/505A61K2039/552Y02A50/30
Inventor 殷玉和吴丛梅李希辰赫玉芳李雪石晶雷欢赵林烨
Owner CHANGCHUN UNIV OF TECH
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